Ultra-Rare Cell In Vivo Flow Cytometry

体内超稀有细胞流式细胞术

基本信息

批准号：
8885325
负责人：
Mark Jonathan Niedre
金额：
$ 34.81万
依托单位：
NORTHEASTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-15 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8885325
关键词：
AMD3100 Adhesions Affect Algorithms Animals Area Biology Biomedical Research Blood Blood Circulation Blood Vessels Blood Volume Blood flow Blood specimen Bone Marrow Cell Count Cell Survival Cells Clinical Trials Collection Combined Modality Therapy Communities Coupled Detection Diffuse Disease Elements Erythrocytes Exploratory/Developmental Grant for Diagnostic Cancer Imaging Fiber Flow Cytometry Fluorescence Frequencies Goals Hematologic Neoplasms Immunology Individual Label Lasers Lead Light Limb structure Malignant Neoplasms Methods Microscopic Molecular Multiple Myeloma Mus National Heart, Lung, and Blood Institute Neoplasm Metastasis Optics Organ Transplantation Outcome Patients Photons Population Reproductive Immunology Reproductive Medicine Research Residual Neoplasm Residual state Resistance SCID Mice Sampling Scanning Technology Testing Tissues Transplantation Work chemotherapy design detector efficacy testing image processing in vivo instrument miniaturize next generation novel novel therapeutics peripheral blood phantom model public health relevance research study signal processing stem cell therapy tool treatment strategy

项目摘要

DESCRIPTION: There are many areas of biomedical research where the study of rare circulating cells is important. Examples include cancer metastasis, hematological malignancies, organ transplant biology, immunology, and reproductive medicine and stem-cell therapies. In this project we will develop a new high-throughput optical scanner with unprecedented capabilities for studying rare circulating cells in small animals in vivo. Circulating cells are normally quantified by drawing small blood samples which are purified and analyzed with hemocytometry or flow cytometry. However, it is known that handling and purifying blood samples can affect cell viability, and that rare cells can escape detection due to the small sampling volume. More recently, `in vivo flow cytometry' (IVFC) methods have been developed that allow enumeration of cells without drawing samples. While extremely useful, these generally rely on interrogation of microscopic blood vessels with small flow rates, so that rare cells are undetectable. New high-sensitivity and high-accuracy tools for studying circulating cells are therefore greatly needed by the research community. In this proposal we will develop a miniaturized optical scanner that will use diffuse photons to interrogate circulating blood in the limb of a mouse. The technology - termed "ultra-rare cell IVFC" (UR-IVFC) - will employ a number of unique design elements including, i) multiple tomographic optical rings with fiber-coupled lasers and fluorescence detectors, ii) efficient geometric light collection, iii) frequency encoded lasers and detector channels, and, iv) advanced signal processing algorithms for accurate counting and tracking of cells. In combination, UR-IVFC will allow single-cell sensitivity in a 10 minute scan and with a false alarm rate less than 0.001 per minute. We anticipate that the unique capabilities of UR-IVFC will have immediate impact in many research fields. We will first use UR-IVFC to study treatment of multiple myeloma (MM), a hematological malignancy for which there is currently no cure. We will study "cell mobilization therapy", which is an emerging treatment strategy for MM. MM cells are chemically forced from the protective bone marrow niche into circulation where they are vulnerable to chemotherapy. We will use UR-IVFC to study whether specific clonal sub-populations of MM cells are resistant to mobilization, and we will test the efficacy of mobilizing agents for minimal residual disease (MRD). In addition to illuminating molecular mechanisms of MM biology and treatment resistance, these studies could yield new therapeutic strategies for patients suffering from MM. Moreover they are extremely difficult (or outright infeasible) to perform with existing technology.

描述：在生物医学研究的许多领域，稀有循环细胞的研究都很重要，例如癌症转移、血液恶性肿瘤、器官移植生物学、免疫学、生殖医学和干细胞疗法。具有前所未有的能力的高通量光学扫描仪，用于研究小动物体内的稀有循环细胞。通常通过抽取小量血液样本来定量这些血液样本，并用血细胞计数法或流式细胞术进行分析。处理和纯化血液样本会影响细胞活力，并且由于采样量小，稀有细胞可能会逃避检测。最近，已经开发出“体内流式细胞术”（IVFC）方法，无需抽取样本即可进行细胞计数。虽然非常有用，但它们通常依赖于以小流速探查显微血管，因此无法检测到用于研究循环细胞的新的高灵敏度和高精度工具。因此，研究界非常需要这种技术，在这项提案中，我们将开发一种小型光学扫描仪，该扫描仪将使用漫射光子来检查小鼠肢体中的循环血液，该技术被称为“超稀有细胞 IVFC”（UR-IVFC）。 ) - 将采用许多独特的设计元素，包括，i) 带有光纤耦合激光器和荧光探测器的多个断层光学环，ii) 高效的几何光收集，iii) 频率编码激光和检测器通道，以及，iv) 用于精确计数和跟踪细胞的先进信号处理算法，结合起来，UR-IVFC 将实现单细胞灵敏度。扫描时间为 10 分钟，误报率低于每分钟 0.001。我们预计 UR-IVFC 的独特功能将在许多研究领域产生直接影响。我们将首先使用 UR-IVFC 来研究多发性骨髓瘤的治疗。 MM），一种目前无法治愈的血液恶性肿瘤，我们将研究“细胞动员疗法”，这是一种新兴的治疗策略，用于将 MM 细胞从保护性骨髓化学环境中强制转移。我们将使用 UR-IVFC 来研究 MM 细胞的特定克隆亚群是否对动员有抵抗力，并且我们将进行测试。除了阐明 MM 生物学和治疗耐药性的分子机制外，这些研究还可以为 MM 患者提供新的治疗策略。使用现有技术执行。